Iodogen法^(125)I标记单克隆抗体10D9

Labeling monoclonal antibody 10D9 with ^(125)I using the Iodogen method

采用Iodogen法,选择最佳标记条件进行125I标记单克隆抗体10D9。标记产物用SephadexG-50分离纯化,三氯醋酸(TCA)法测定标记率和放化纯,并对标记物的免疫活性及稳定性进行分析。结果显示,125I标记10D9的最佳条件为Iodogen用量10μg、10D9用量20μg、加入Na125I22.2MBq、室温反应8min,125I-10D9的标记率为(84.10±3.18)%,放化纯为(98.49±1.14)%,比活度为933.51MBq/mg;125I-10D9与Daudi细胞的特异性结合率为(23.21±1.14)%;标记物加到含1%BSA的PBS中放置3W后放化纯度仍>90%。结果表明:Iodogen法125I标记10D9标记率和放化纯高,方法简便,标记物的稳定性好且能较好地保持其免疫活性。

To establish the method radiolabeled monoclonal antibody 10D9 against human costimulatory molecules CD80 with ^125I, 10D9 was labeled with ^125I using the Iodogen method at room temperature. The labeling efficiency and radiochemical purity were tested with TCA, and the immune activity and stability of the labeling product were analyzed. The optimum labeling condition was 10 μg Iodogen, 20 μg 10D9 and 22.2 MBq Na^125I. The reaction time was 8 min at room temperature. The labeled yield was （84.10±3.18）%, and the radiochemical purity was （98.49±1.14）%. The radio-activity of ^125I-10D9 was 933.51 MBq/mg. The specific binding efficiency of ^125I-10D9 with Daudi cell was （23.21±1.14）%. The radiochemical purity of ^125I-10D9 was over 90% after three weeks of mixing with PBS. In conclusion, the method is simple and easy to perform. The labeling efficiency and radiochemical purity is high. The stability of ^125I-10D9 is fine in vitro and ^125I-10D9 keeps its immune activity after Iodogen labeling.